α-葡聚糖酶在毕赤酵母中的组成型表达  

Constitutive Expression of Dextranase in Pichia pastoris

作　　者：梁达奉[1,2] 黄曾慰[2] 曾练强[2] 蚁细苗[2] 李雨虹[2] 余林[1] 

机构地区：[1]广东工业大学轻工化工学院,广东广州510006 [2]广州甘蔗糖业研究所广东省甘蔗改良与生物炼制重点实验室,广东广州510316

出　　处：《华南理工大学学报（自然科学版）》2012年第5期96-100,共5页Journal of South China University of Technology(Natural Science Edition)

基　　金：科技部科研院所技术开发研究专项资金资助项目(NCSTE-2007-JKZX-023);国家科技支撑计划项目(2011BAE16B03)

摘　　要：为避免α-葡聚糖造成的制糖过程中的糖分损失、制炼难度增加以及糖产品质量下降,利用α-葡聚糖酶分解而直接去除α-葡聚糖是目前最佳的选择.文中扩增了α-葡聚糖酶基因dex,构建组成型表达载体pGAPZαA-dex;以毕赤酵母KM71H为受体菌,将表达载体整合进入受体菌基因组,构建了组成型表达α-葡聚糖酶的毕赤酵母工程菌.摇瓶发酵120h,α-葡聚糖酶酶活达到60.4U/mL,从而实现了α-葡聚糖酶在毕赤酵母中的组成型表达,使发酵过程无需使用甲醇进行诱导,提高了生产和使用安全性.Enzymolysis of using dextranase is the best way to the removal of dextrans that result in sugar loss,manufacture difficulty and quality decline during the sugar manufacture.In this paper,dextranase gene dex was cloned into a yeast constitutive expression vector pGAPZαA,and the recombinant plasmid pGAPZαA-dex was transformed into Pichia pastoris KM71H to construct an engineering strain constitutively expressing dextranase.After the cultivation for 120 h in a shake flask,dextranase with an activity of 60.4 U/mL was successfully obtained,thus implementing the constitutive expression of dextranase in Pichia pastoris.The proposed method avoids the use of me-thanol during the fermentation,which helps to improve the production and use safety during the sugar manufacture.

关 键 词：制糖 α-葡聚糖酶 组成型表达 毕赤酵母 发酵 

分 类 号：Q786[生物学—分子生物学]